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Developing a time-dynamic, label-free assay for apoptosis based on optical Gabor filtering

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TitleInfo
Title
Developing a time-dynamic, label-free assay for apoptosis based on optical Gabor filtering
Name (type = personal)
NamePart (type = family)
Pasternack
NamePart (type = given)
Robert M
NamePart (type = date)
1985-
DisplayForm
Robert Pasternack
Role
RoleTerm (authority = RULIB)
author
Name (type = personal)
NamePart (type = family)
Boustany
NamePart (type = given)
Nada N
DisplayForm
Nada N Boustany
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
chair
Name (type = personal)
NamePart (type = family)
Yarmush
NamePart (type = given)
Martin l
DisplayForm
Martin l Yarmush
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
internal member
Name (type = personal)
NamePart (type = family)
Metaxas
NamePart (type = given)
Dimitris N
DisplayForm
Dimitris N Metaxas
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
internal member
Name (type = personal)
NamePart (type = family)
White
NamePart (type = given)
Eileen P
DisplayForm
Eileen P White
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
outside member
Name (type = corporate)
NamePart
Rutgers University
Role
RoleTerm (authority = RULIB)
degree grantor
Name (type = corporate)
NamePart
Graduate School - New Brunswick
Role
RoleTerm (authority = RULIB)
school
TypeOfResource
Text
Genre (authority = marcgt)
theses
OriginInfo
DateCreated (qualifier = exact)
2011
DateOther (qualifier = exact); (type = degree)
2011-05
Place
PlaceTerm (type = code)
xx
Language
LanguageTerm (authority = ISO639-2b); (type = code)
eng
Abstract (type = abstract)
Apoptosis is a fundamental process in the homeostasis and development of mammals, dysfunction of its regulation is implicated in human disease, and it is of growing interest as a possible therapeutic target (especially in cancer). An apoptosis assay that is sensitive, quantitative, and label-free would therefore be useful in cancer drug discovery by offering a direct, time-dynamic in-situ assessment of apoptotic response to drug candidates. This thesis documents the development of optical Gabor filtering into an apoptosis assay. Optical Gabor filtering was developed as an improvement to optical scatter imaging (OSI) capable of optical scatter-based detection of object size and aspect ratio without the need for a predictive scatter model or extrinsic labeling. Two versions of the optical Gabor filtering instrument (LCD-based and DMD-based). By applying a series of Gabor filters with varying spatial frequencies and orientations, a series of filtered images can be generated and processed to extract morphological parameters of interest. The parameters of size, orientedness and aspect ratio are developed and tested using polystyrene microspheres, marine diatoms and collagen gelation, respectively. Orientedness and aspect ratio are employed as morphometric parameters to study dynamic morphological alterations in apoptosis. Aspect ratio measurements in apoptotic iBMK cells show a differential aspect ratio response to apoptosis induction by bax/bak expressing and bax/bak knockout cell variants. To determine the biological source of these variations, multimodal image acquisition of fluorescently-labeled mitochondria alongside Gabor filtered data is used in apoptotic endothelial cells. Results of this study show a drop in orientedness 1-2 h post-STS treatment in mitochondria-rich regions (reconnoitered by fluorescence) concomitant with mitochondrial fragmentation but absent elsewhere. This dynamic is similar but not identical to that observed from iBMK cells. OSI imaging of apoptotic BAEC also revealed that a previously reported drop in optical scatter image ratio in the first hour of apoptosis is also associated with mitochondria-rich regions. In the future, the differences between optical responses of BAEC and iBMK apoptotic cells will be elucidated with fluorescent tracking of mitochondria-rich areas in iBMK. Classification and other computational methods could also be employed to directly evaluate the optical Gabor-filtered data in the future.
Subject (authority = RUETD)
Topic
Biomedical Engineering
RelatedItem (type = host)
TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier (type = RULIB)
ETD
Identifier
ETD_3261
PhysicalDescription
Form (authority = gmd)
electronic resource
InternetMediaType
application/pdf
InternetMediaType
text/xml
Extent
xv, 156 p. : ill.
Note (type = degree)
Ph.D.
Note (type = bibliography)
Includes bibliographical references
Note (type = vita)
Includes vita
Note (type = statement of responsibility)
by Robert M Pasternack
Subject (authority = ETD-LCSH)
Topic
Apoptosis
Subject (authority = ETD-LCSH)
Topic
Optoelectronic devices
Identifier (type = hdl)
http://hdl.rutgers.edu/1782.1/rucore10001600001.ETD.000061437
RelatedItem (type = host)
TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
Identifier (type = local)
rucore19991600001
Location
PhysicalLocation (authority = marcorg); (displayLabel = Rutgers, The State University of New Jersey)
NjNbRU
Identifier (type = doi)
doi:10.7282/T36M3651
Genre (authority = ExL-Esploro)
ETD doctoral
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Rights

RightsDeclaration (ID = rulibRdec0006)
The author owns the copyright to this work.
RightsHolder (type = personal)
Name
FamilyName
Pasternack
GivenName
Robert
Role
Copyright Holder
RightsEvent
Type
Permission or license
DateTime (encoding = w3cdtf); (qualifier = exact); (point = start)
2011-04-13 11:34:24
AssociatedEntity
Name
Robert Pasternack
Role
Copyright holder
Affiliation
Rutgers University. Graduate School - New Brunswick
AssociatedObject
Type
License
Name
Author Agreement License
Detail
I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
Copyright
Status
Copyright protected
Availability
Status
Open
Reason
Permission or license
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